Kotani M, Yamamura Y, Tamatani T, Kitamura F, Miyasaka M
Department of Immunology, Tokyo Metropolitan Institute of Medical Science, Japan.
J Immunol Methods. 1993 Jan 4;157(1-2):241-52. doi: 10.1016/0022-1759(93)90093-m.
We describe in this report the production and characterization of monoclonal antibodies (mAb) to the rabbit homologues of CD4, CD5 and CD11a antigens, and their use for phenotypic analysis of rabbit lymphoid cell lines. All the mAbs were produced by immunizing mice with rabbit thymocytes. mAb KEN-4 apparently identified rabbit CD4, precipitated two bands of 42 and 50 kDa under reducing and non-reducing conditions and markedly inhibited allo-MLR. The distribution of antigen-positive cells were restricted to the thymus and classical T-dependent areas in peripheral lymphoid tissues. mAb KEN-5 apparently identified rabbit CD5, precipitated a single polypeptide of 67 kDa similar to other anti-CD5 mAb in the human and mouse. The use of this mAb revealed that CD5+ B cells were infrequent in this species. mAb KEN-11 apparently identified rabbit CD11a and precipitated a heterodimer of 150/95 kDa by selectively recognizing the 150 kDa moiety. It blocked cation-dependent aggregation of phorbol ester-induced rabbit Con A blasts and also allo-MLR in a similar manner to other anti-CD11a mAb in various animal species. Phenotypic examination of HTLV-1 transformed rabbit lymphoid cell lines using these mAb clearly indicated that most of them were CD4+, CD5+ and CD11a+, and hence derived from CD4+ T cells. These mAb will be useful tools for the study of the cellular immune system in the rabbit.
我们在本报告中描述了针对兔源CD4、CD5和CD11a抗原同系物的单克隆抗体（mAb）的制备及特性鉴定，以及它们在兔淋巴细胞系表型分析中的应用。所有单克隆抗体均通过用兔胸腺细胞免疫小鼠制备。单克隆抗体KEN-4明显识别兔CD4，在还原和非还原条件下沉淀出两条分别为42 kDa和50 kDa的条带，并显著抑制同种异体混合淋巴细胞反应（allo-MLR）。抗原阳性细胞的分布局限于胸腺和外周淋巴组织中的经典T细胞依赖区。单克隆抗体KEN-5明显识别兔CD5，沉淀出一条67 kDa的单一多肽，类似于人和小鼠中的其他抗CD5单克隆抗体。使用该单克隆抗体发现，该物种中CD5+B细胞很少见。单克隆抗体KEN-11明显识别兔CD11a，并通过选择性识别150 kDa部分沉淀出150/95 kDa的异二聚体。它阻断佛波酯诱导的兔刀豆蛋白A刺激细胞的阳离子依赖性聚集，也以与各种动物物种中的其他抗CD11a单克隆抗体类似的方式阻断同种异体混合淋巴细胞反应。使用这些单克隆抗体对人嗜T细胞病毒1型（HTLV-1）转化的兔淋巴细胞系进行表型检测清楚地表明，它们中的大多数是CD4+、CD-5+和CD11a+，因此源自CD4+T细胞。这些单克隆抗体将成为研究兔细胞免疫系统的有用工具。
